A robust method for the rapid generation of recombinant Zika virus expressing the GFP reporter gene

- The reverse genetic method ISA is a convenient method for rapid production of ZIKV molecular clones.
- Molecular clones are reliable tools for investigation on ZIKV viral determinants of human disease.
- The ISA method allows the rapid production of recombinant ZIKVGFP expressing the GFP reporter gene.
- ZIKVGFP is suitable for dynamics study on viral replication in host cells from mosquito to human.
- ZIKVGFP is suitable to test antiviral activity of inhibitors as well as of neutralizing antibodies.

Zika virus (ZIKV) infection is a major public health problem with severe human congenital and neurological anomalies. The screening of anti-ZIKV compounds and neutralizing antibodies needs reliable and rapid virus-based assays. Here, we described a convenient method leading to the rapid production of molecular clones of ZIKV. To generate a molecular clone of ZIKV strain MR766NIID, the viral genome was directly assembled into Vero cells after introduction of four overlapping synthetic fragments that cover the full-length genomic RNA sequence. Such strategy has allowed the production of a recombinant ZIKV expressing the GFP reporter gene that is stable over two culturing rounds on Vero cells. Our data demonstrate that the ZIKV reporter virus is a very reliable GFP-based tool for analyzing viral growth and measuring the neutralizing antibody as well as rapid screening of antiviral effect of different classes of inhibitors.